LC ESI QOTOF METHOD DEVELOPMENT AND NEW INSTRUMENTATION EVALUATION

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 我们的实验室使用两台Applied Biosystems API QSTAR（tm）Pulsar i QqTOF质谱仪和一台配备沃茨Acuity LC的Agilent chip-LC Q-TOF质谱仪，通过ESI和MALDI对从合作研究项目中获得的复杂生物样品进行表征。目前，我们的重点是建立常规毛细管LC-IDA-MS/MS用于蛋白质组学，糖组学和脂质组学应用，以及表征提交给MS资源的各种生物样品。自动化IDA MS/MS功能的使用正在针对典型的蛋白质代谢物、脂质和碳水化合物进行不断优化，并且已经生成了批参数文件，以便于实验室内的成员使用。处理脚本已经过优化，可在工作站上用于数据分析。使用较小直径的色谱柱组件可显著提高灵敏度，从而能够分析低浓度或有限数量的样品。LC系统的优化之后是方法开发研究，以补充MS资源内进行的当前研究。正在为糖组学、脂质组学和蛋白质组学研究开发方法，包括磷酸化分析。将Advion TriVersa机器人添加到Q-Star系统中需要进行大量工作以找到最佳条件，并与Advion和Applied Biosystems密切互动。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Our laboratory uses two Applied Biosystems API QSTAR(tm) Pulsar i QqTOF mass spectrometers with Waters Acuity LCs and an Agilent chip-LC Q-TOF mass spectrometer for the characterization of complex biological samples obtained from collaborative research projects by ESI and MALDI. Currently, our focus is on establishing routine capillary LC-IDA-MS/MS for proteomics, glycomics and lipidomics applications, in addition to characterization of the large variety of biological samples that are submitted to the MS Resource. Use of automated IDA MS/MS functions are continuously being optimized for typical protein digests, lipids and carbohydrates, and batch parameter files have been generated for ease of use by the members within the laboratory. Processing scripts have been optimized for use on workstations for data analysis. The use of the smaller diameter column assemblies offers a significant increase in sensitivity, enabling samples of low concentrations or limited quantity to be analyzed. Optimization of the LC systems is followed by method development studies to complement the current research undertaken within the MS resource. Method development is being pursued for glycomics, lipidomics and proteomics-based studies, including phosphorylation analysis. Addition of the Advion TriVersa robot to the Q-Star system has necessitated much work to find optimum conditions, and close interaction with both Advion and Applied Biosystems.